! PROJECT SUMMARY/ABSTRACT Expanding our understanding of the genetic contributions and etiologies of birth defects and childhood cancer will have a significant and direct impact on those affected by those conditions and contribute significantly to research in related adult conditions. This proposal continues the HudsonAlpha-St. Jude Genome Sequencing Center (HASJ-GSC) and expands its capabilities. The HASJ-GSC will provide an efficient and experienced genomics resource to generate and analyze high-quality sequence and variant data from the largest number of pediatric cancer and birth defect cohorts possible using the funds available over a three-year period. The innovation, capabilities, and experience of the HASJ-GSC will produce data for the Gabriella Miller Kids First Research Program that will be accessible and available to the research community and leveraged to its maximal impact for years to come. A robust cost reduction plan allows for an increase in the number of cases that can be sequenced and analyzed per dollar per year. The HASJ-GSC will generate exceptional quality whole-genome sequence and variant data for all samples and for the pediatric cancer samples, RNASeq and whole-exome data. The combined genome, exome and RNASeq will provide as much resolution as possible to understand the genetic and functional genomic changes observed in pediatric cancer. The HASJ-GSC will also provide support for a comprehensive collection of additional methodologies such as long read, lined-long read and RNAseq. These will expand the resolution and type of investigation that can be supported for X01 investigators. The HASJ-GSC will also provide a reliable and efficient data storage and data access capability that provides fast, reliable and efficient data access, sharing and reporting for the X01 projects and samples. Finally, the HASJ-GSC will support efficient submission of sequence and variant data to the Gabriella Miller Kids First Data Resource Center and help facilitate submission to the appropriate public databases. !